Journal: International Journal of Molecular Sciences

Article Title: Mating Increases CHST10 Activity in Rat Oviductal Mucosa to Induce the Synthesis of HNK-1 Glycoproteins: Possible Role in Sperm–Oviduct Interactions

doi: 10.3390/ijms26073309

Figure Lengend Snippet: Mating increases the carbohydrate sulfotransferase 10 (CHST10) protein level in the oviductal mucosa of rats. Western blot of samples obtained from the oviductal mucosa of unmated (UM1-4) and mated (M1-4) rats. Biological replicates for each group, n = 4. The bar graph represents the intensities of the three CHST10 isoforms detected by the Western blot and normalized to that of tubulin β class I (TUBB). Interquartile range (IQR): Interquartile range. * p < 0.05, indicates significant differences.

Article Snippet: We also evaluated the substrate specificity of human recombinant CHST10 (R&D Systems, cat# 6140-ST) and compared the activity of this enzyme to the sulfotransferase activity detected in oviductal mucosa protein extracts.

Techniques: Western Blot

Journal: International Journal of Molecular Sciences

Article Title: Mating Increases CHST10 Activity in Rat Oviductal Mucosa to Induce the Synthesis of HNK-1 Glycoproteins: Possible Role in Sperm–Oviduct Interactions

doi: 10.3390/ijms26073309

Figure Lengend Snippet: Expression of CHST10 in the rat oviduct. CHST10 was detected via immunofluorescence in oviductal sections obtained from unmated and mated rats. The panels show merged red and blue fluorescence. CHST10: red; and nuclear staining: blue. ( First row ) ampulla; ( second row ) isthmus; and ( third row ) utero-tubal junction (UTJ). The panels in the left column are the negative controls. White arrowheads: immune cells in negative controls; red arrowheads: CHST10 signal resembling the Golgi apparatus location; red arrows: CHST10 in the epithelium cytoplasm. L, Lumen; E, mucosa layer; M, muscular layer. Original magnification, 200× (scale bar, 75 μm). Only the right panels display the image at 630× magnification (scale bar, 25 μm). Images are representative of three independent experiments.

Article Snippet: We also evaluated the substrate specificity of human recombinant CHST10 (R&D Systems, cat# 6140-ST) and compared the activity of this enzyme to the sulfotransferase activity detected in oviductal mucosa protein extracts.

Techniques: Expressing, Immunofluorescence, Fluorescence, Staining

Journal: International Journal of Molecular Sciences

Article Title: Mating Increases CHST10 Activity in Rat Oviductal Mucosa to Induce the Synthesis of HNK-1 Glycoproteins: Possible Role in Sperm–Oviduct Interactions

doi: 10.3390/ijms26073309

Figure Lengend Snippet: Mating increases CHST10 activity in the oviductal mucosa of rats. ( A ) Michaelis-Menten kinetics of human recombinant CHST10 to evaluate specific acceptor substrates. PGA: phenolphthalein glucuronic acid; E 2 3G: 17β-estradiol 3-[β-D-glucuronide]. Biological replicates for each group, n = 3. ( B ) Pattern of acceptor substrate preferences for CHST10 ( left ) and total protein from the oviductal mucosa of unmated rats ( right ). CSA: chondroitin sulfate A; E 2 : 17β-estradiol; E 2 17G: 17β-estradiol 17-[β-D-glucuronide]; DMSO: dimethyl sulfoxide. Biological replicates for each group, n = 3. ( C ) Plots of sulfotransferase activity measured in the oviductal mucosa of unmated and mated rats using PGA and E 2 3G as acceptor substrates. Biological replicates for each group, n = 8. *** p < 0.001, indicates significant differences.

Article Snippet: We also evaluated the substrate specificity of human recombinant CHST10 (R&D Systems, cat# 6140-ST) and compared the activity of this enzyme to the sulfotransferase activity detected in oviductal mucosa protein extracts.

Techniques: Activity Assay, Recombinant

Journal: International Journal of Molecular Sciences

Article Title: Mating Increases CHST10 Activity in Rat Oviductal Mucosa to Induce the Synthesis of HNK-1 Glycoproteins: Possible Role in Sperm–Oviduct Interactions

doi: 10.3390/ijms26073309

Figure Lengend Snippet: Vaginocervical Stimulation (VCS) increases the level of Chst10 in the oviductal mucosa of rats. The graphs show the RT-qPCR results for Chst10 normalized to the expression of reference genes. The scatter plots represent the normalized individual data points after 2 −ΔCq transformation for each individual sample. ( A ) Oviductal mucosa obtained 3 h after intrauterine injection of 0.1 mL of saline (0.9% w / v NaCl), a suspension of sperm in saline (10 million cells), or diluted seminal vesicle fluid (SVF, 1:1 dilution) into unmated rats. Biological replicates for each group, n = 5. ( B ) Oviductal mucosa obtained from unmated rats 3 h after VCS or sham stimulation. Biological replicates for each group, n = 5. ** p < 0.01, indicates significant differences.

Article Snippet: We also evaluated the substrate specificity of human recombinant CHST10 (R&D Systems, cat# 6140-ST) and compared the activity of this enzyme to the sulfotransferase activity detected in oviductal mucosa protein extracts.

Techniques: Quantitative RT-PCR, Expressing, Transformation Assay, Injection, Saline, Suspension